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Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

Bpu10I

5'...C C↓T N A G C...3'
3'...G G A N T↑C G...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Unique buffer for 100% activity Optimal incubation at 37°C Thermal inactivation at 80°C in 20 min Recombinant enzyme LO certified

The Bpu10I restriction enzyme recognizes CCTNAGC(-5/-2)^ sites and cuts best at 37°C in its own unique buffer
  
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Lambda DNA digested with Bpu10I

Lambda DNA digested with Bpu10I, 0.7% agarose, 19 cleavage sites

Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

A large excess of enzyme (> 4 U/1 µg DNA) may result in incomplete DNA cleavage. Therefore, we recommend increasing the incubation time instead of using an excess of Bpu10I. Low salt, high glycerol (> 5%) concentrations, pH > 7.0, or a large excess of enzyme may result in star activity. For cleavage with Bpu10I, at least two copies of its recognition sequence are required.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsBpu1102I, DdeI, Eco81I.
Conditions for 100% Activity
  • 1x Buffer Bpu10I: 10 mM Bis-Tris Propane-HCl (pH 6.5 at 37°C), 10 mM MgCl2, 100 mM KCl, and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBpu10I is supplied in:
10 mM Tris-HCl (pH 7.5 at 25°C), 200 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X		 G (green)
1X		 O (orange)
1X		 R (red)
1X		 Tango (yellow)
1X / 2X
Bpu10I Buffer (Unique) 37°C 0-20 20-50† 50-100† 100† 50-100† 100† 1X† or 2X†

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

Methylation type Sequence Cleavage effect
CpG 5'...CCTNAGm5C G...3'
3'...GGANTC Gm5C...5'		 No effect
EcoBI (TGA(N)8TGCT) 5'...CCTGm6AGC(N)6 TGCT...3'
3'...GGAC TCG(N)6m6ACGA...5'		 No effect

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
19 7 4 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 5 3

New sites generated by ligation

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
CC^TCAGCCCTCATCAGC
  • [MnlI/FastDigest MnlI]